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A kind of enzyme-labeled secondary antibody complex and preparation method thereof

An enzyme-labeled secondary antibody and complex technology, applied in the field of immunoassay, can solve the problems of limited number of enzymes and antibodies, insufficient sensitivity, low penetration, etc., to reduce hydrodynamic volume, improve sensitivity, and improve penetration Penetration effect

Active Publication Date: 2021-04-02
河南赛诺特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is not perfect, and its main technical defects are: (1) the method adopts a dextran chain polymer backbone, which has a loose structure and a huge hydrodynamic volume, thereby causing great steric hindrance, making the polymer less permeable to the nuclear membrane
Therefore, the performance of this polymer for the detection of various types of antigens is uneven, especially in the detection of low-expressed antigens in some nuclei; (2) the method is to directly label enzymes and intact antibodies On the backbone, due to the limited surface area on the backbone, the number of enzymes and antibodies that can be labeled is limited, so the sensitivity cannot be further improved

Method used

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  • A kind of enzyme-labeled secondary antibody complex and preparation method thereof
  • A kind of enzyme-labeled secondary antibody complex and preparation method thereof
  • A kind of enzyme-labeled secondary antibody complex and preparation method thereof

Examples

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Effect test

Embodiment 1

[0065]The enzyme-labeled secondary antimethyrridate Boltorn, including hyperbranched aminocyreneTM H40 and horseradish peroxidase, horseradish peroxidase is attached to superblyminosylated polydimethyl hydroxypropionate BoltornTM On the H40, the macama peroxidase was attached to the u.acely-ethyl ester, and the lambs were attached to the sour diethylate.

[0066]Super branched aminocyanocyrene hydroxypropionate BoltornTM At its branched end and the middle partial support has an amino group that can be coupled to the coupling agent; aminocylated boltornTM Structure of series products, such asFigure 4 Indicated.

[0067]The principle of immunoassay of the enzyme-labeled secondary anti-composite of this embodiment is shown.Figure 5 As shown, the enzyme and antibody fragments are stacked in the surface of the superblyminatous polydimethylene hydroxypropylene skeleton.

Embodiment 2

[0069]The enzyme-labeled secondary antimethyrridate Boltorn, including hyperbranched aminocyreneTM H40 and horseradish peroxidase, horseradish peroxidase is attached to superblyminosylated polydimethyl hydroxypropionate BoltornTM On the H40, a sheep anti-rabbit antibody IgG-Fab 'was attached to the sour diendoxide.

Embodiment 3

[0071]The enzyme-labeled secondary antimethyrridate Boltorn, including hyperbranched aminocyreneTM H40 and horseradish peroxidase, horseradish peroxidase is attached to superblyminosylated polydimethyl hydroxypropionate BoltornTM On the H40, a horseradish peroxidase was attached to the othyl ester (ABCAM, SKU: AB98634).

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Abstract

The invention relates to an enzyme-labeled secondary antibody compound and a preparation method thereof, and belongs to the technical field of immunoassay. The enzyme-labeled secondary antibody compound comprises a polymer skeleton and a labeling enzyme. The labeling enzyme is connected to the polymer skeleton through a coupling agent, and a secondary antibody is connected to the labeling enzyme through a coupling agent; wherein the polymer skeleton is provided with a branched chain end part and a middle branch structure connected with the branched chain end part, and the labeling enzyme is connected to the branched chain end part and the middle branch structure of the polymer skeleton through a coupling agent; and the polymer skeleton is a hyper-branched polymer skeleton. The branched chain end part and the middle branch structure of the polymer skeleton can be connected with a coupling agent, so that the sensitivity of the enzyme-labeled secondary antibody compound is favorably improved, and the accumulation mode of enzyme and antibody is favorable for improving the polymerization degree of enzyme and antibody in the polymer, so that the sensitivity of the enzyme-labeled secondary antibody compound in immunoassay is greatly improved.

Description

Technical field[0001]The present invention relates to an enzyme-laid secondary anti-complex and a preparation method thereof, belonging to the technical field of immunoassay.Background technique[0002]Immunoassay, that is, the method of detecting samples of protein biomarkers by "antibody-antigen specific binding". Immunoassays can be widely used in the qualitative and quantitative analysis of various biological samples due to their height sensitivity and specificity. Methods of immunoassay include: Enzyme-linked immunization (ELISA), Western-blot, immunohistochemistry, lateral flow analysis method, protein array, cellular immunity ( Immunocytochemistry, etc. Wherein "HRP secondary antibody complex" is widely used to detect biomarkers in the field of immunoassays, including its applications immunohistochemistry, enzyme immunoassay, immunoblotting, and other immune cells, "HRP secondary antibody complex" the principle of the sample is identified by a specific antibody complex in the t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535
CPCG01N33/535
Inventor 刘畅梁永波牛银银刘玲玲齐华
Owner 河南赛诺特生物技术有限公司
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